猪GRP78基因克隆、生物信息学分析及组织表达谱研究

doi:10.16431/j.cnki.1671-7236.2018.12.002

Abstract

Abstract:

This study was aimed to clone and analyze the CDS region of porcine glucose-regulated protein 78 (GRP78) gene using bioinformatics,and investigate the mRNA expression profile of GRP78 gene.Primers were designed according the predicted sequence of porcine GRP78 gene (accession No.:XM 001927795.6) in GenBank.The CDS region of porcine GRP78 gene was cloned by PCR amplification and sequencing.The physical and chemical property,transmembrane region,signal peptide,hydrophobicity,conserved domains,the secondary and tertiary structures were analyzed,and the phylogenetic tree of GRP78 gene was constructed by online prediction softwares.Finally,Real-time PCR was used to detect the tissue expression of porcine GRP78 gene.The results showed that the CDS of GRP78 gene was 1 965 bp,encoding 654 amino acids.Bioinformatics analysis showed that the molecular weight,theoretical electrical point (pI),half-life and the extinction coefficient of water solution at 280 nm of GRP78 were 72.3 ku,5.06,30 h,and 30 495,respectively.The peptide chain N-terminal of GRP78 was M (Met).The unstsble coefficient of GRP78 was 32.39,indicating that it belonged to the stable protein.The fat coefficient of GRP78 protein was 85.40,and the total average hydrophobic index was -0.496.GRP78 did not have transmembrane domain but it had one signal peptide,therefore,this protein belonged to the secreted protein.The secondary structure analysis of GRP78 showed that the percent of amino acids of α-helix,extended chain,β-turn and random coil were 40.06%,20.49%,8.10% and 31.35%,respectively.Homology analysis results showed that the nucleotide homology of GRP78 gene with human(NM_005347.4),mouse(NM_001163434.1),rat(NM_013083.2),goat (XM_005687138.3) and cattle (NM_001075148.1) were 93%,91%,90%,95% and 95%,whereas the amino acid homology were 99%,98%,98%,99% and 99%,respectively.These results indicated that GRP78 gene was highly conserved.Real-time PCR results showed that GRP78 gene was expressed in heart,liver,spleen,lung,kidney,small intestine,stomach,ovary,fallopian tube,breast,cerebellum,cerebrum and pituitary gland,and highly expressed in heart,spleen and lung.This study provided basic materials for further study of the biological function of GRP78 gene.

Key words: porcine; GRP78 gene; clone; bioinformatics analysis; tissue expression

CLC Number:

ZHAO Shiyu, ZHANG Fan, ZHOU Xiaolong, WANG Han, ZHAO Ayong, YANG Songbai. Cloning,Bioinformatics and Tissue Expression Analysis of Porcine GRP78 Gene[J]. , 2018, 45(12): 3317-3326.

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Cloning,Bioinformatics and Tissue Expression Analysis of Porcine GRP78 Gene

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